The document summarizes the process of extracting and isolating a compound from the Chinese herbal medicine Erigeron breviscapus that has depigmenting effects. Materials and methods describe culturing cells and conducting inhibition and isolation assays. Through fractionation of the plant extract, the active compound (2Z,8Z)-Matricaria Acid Methyl Ester was isolated. Results show the compound decreased melanin synthesis and tyrosinase expression in cells, demonstrating its potential as a skin whitening agent for treating hyperpigmentation.

1. Objective: to be able to understand the process of extraction and Isolation
Leonard Guinguino
13-11-23

2. Introduction
• Erigeron breviscapus
- A chinese herbal medicine that has
different pharmacologic effect is studied
and is shown to posses dipegmenting
effect.
- the extract was collected and isolation of
the compound (2Z,8Z)- Matricaria Acid
Methyl Ester was conducted.

3. Materials and Methods
Cell culture and Inhibition
Assay
•DMEM
•PBS
•FBS
•DMSO
•1N NaOH
•MTT solution
•Kojic Acid
•PTU
•Triton X
Isolation of Active
compounds
•Erigeron breviscapus
•Methanol
•Hexane
•Column chromatograph
•Chloroform
•NMR

4. Materials and methods
Isolation of (2Z,8Z)- through fractionation

5. Results
Fig. 1. Effect of Methanol Crude Extract on
Melanin Synthesis in B16F10 Melanoma
Cells
Fig. 4. Effect of (2Z,8Z)-Matricaria Acid
Methyl Ester on Melanin Synthesis in
Melan-a Cells
Fig. 6. Effect of (2Z,8Z)-Matricaria Acid Methyl Ester
on Tyrosinase Expression in Melan-a Cells
•Decreased tyrosinase protein level was found by
western blot analysis.

6. Results
Fig. 5. Effect of (2Z,8Z)-Matricaria Acid Methyl Ester on
(A)Mushroom Tyrosinase,
•No inhibition of in vitro mushroom tyrosinase activity was
observed by the isolated compound.
(B) Cellular Tyrosinase from B16F10
•a significant decrease in cellular tyrosinase activity was
observed in the compound treated cells
(C) Melan-a Cells.
•a significant decrease in cellular tyrosinase activity was observed in the
compound treated cells

7. Conclusion
•
Found that this plant contains a melanin
biosynthesis inhibitor, which was isolated and identified as (2Z,8Z)-matricaria
methyl ester.
• These results showed that (2Z,8Z)-matricaria acid methyl ester isolated
from Erigeron breviscapus can be a promising compound that could be used as
skin whitening agent for treating hyperpigmentation.